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BACKGROUND: Jackfruit seed flour can be used as a cocoa aroma replacer with similar technological properties. The purpose of this study was to investigate the in vivo toxicity and in vitro antioxidant activity of fermented jackfruit seed flour (Fjs) and non‐alkaline cocoa powder (Nac). RESULTS: Fjs and Nac extracts (Fjs‐E and Nac‐E) were produced and submitted to in vitro gastrointestinal digestion producing digested fractions named Fjs‐D and Nac‐D, respectively. Nac‐E showed over two‐fold higher oxygen radical absorbance capacity (ORAC) than Fjs‐E. However, after simulated gastrointestinal digestion (in vitro), there were no significant differences between Nac‐D and Fjs‐D (P < 0.01). Similarly, the cellular antioxidant activity (CAA) of Nac‐D and Fjs‐D was not significantly different (P < 0.01). The anti‐inflammatory assay in transgenic RAW 264.7 murine macrophages showed that Fjs‐E did not affect cell viability up to 300 μg mL−1 (P > 0.05) and reduced by 15% the release of TNF‐α (P < 0.05). Fjs‐D did not affect cell viability up to 300 μg mL−1 (P > 0.05) and showed 58% reduction of NF‐κB activation (P < 0.05), with no effects on TNF‐α levels. Treatment with Nac‐E up to 300 μg mL−1 did not decrease cell viability (P > 0.05) and reduced the release of TNF‐α levels by 34% and 66% at 100 and 300 μg mL−1, respectively (P < 0.05). Nac‐D did not reduce the NF‐κB activation or TNF‐α levels at any tested concentration. CONCLUSION: Collectively, these findings indicate that Fjs is a safe and promising functional ingredient with biological activities even after gastrointestinal digestion. © 2023 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

In the present study, a novel water-soluble polysaccharide (DNP−1) was isolated and purified from the root of Duhaldea nervosa via column chromatography. Structural analyses indicated that DNP−1 had a linear backbone consisting of (2→1)-linked β-D- fructofuranosyl residues, ending with a (2→1) bonded α-D-glucopyranose. DNP−1 was a homogeneous polysaccharide with an average molecular weight of 3.7 kDa. Furthermore, the anti-inflammatory activity of DNP−1 was investigated in vitro. The concentration of pro-inflammatory cytokines, including NO, TNF-α, MCP-1, IL-2, and IL-6, in the DNP−1 treatment group was suppressed in LPS-induced RAW 264.7 cells. DNP−1 was able to improve inflammatory injury by inhibiting the secretion of pro-inflammatory cytokines. These investigations into this polysaccharide from the root of Duhaldea nervosa provide a scientific basis for the further development of this plant. The results indicate that this Duhaldea nervosa polysaccharide could be used as a potential natural source for the treatment of inflammatory injury. [ABSTRACT FROM AUTHOR]

Host inflammatory responses are key to protection against injury; however, persistent inflammation is detrimental and contributes to morbidity and mortality. Herein, we demonstrated the anti-inflammatory role of Arteannuin-B (1) and its new spirocyclic-2-isoxazoline derivative JR-9 and their side effects in acute inflammatory condition in vivo using LPS-induced cytokines assay, carrageenan-induced paw edema, acetic acid-induced writhing and tail immersion. The results show that the spirocyclic-2-isoxazoline derivative is a potent anti-inflammatory agent with minimal cell toxicity as compared to Arteannuin-B. In addition, the efficacies of these compounds were also validated by flow cytometric, computational, and histopathological analysis. Our results show that the anti-inflammatory response of JR-9 significantly reduces the ability of mouse macrophages to produce NO, TNF-α, and IL-6 following LPS stimulation. Therefore, JR-9 is a prospective candidate for the development of anti-inflammatory drugs and its molecular mechanism is likely related to the regulation of NF-κB and MAPK signaling pathway. [ABSTRACT FROM AUTHOR]

Antrodia camphorata is an endemic mushroom in Taiwan. This study was designed to screen anti-inflammatory compounds from the methanolic extract of the mycelium of A. camphorata on nitric oxide (NO) production in RAW 264.7 cells induced by polyinosinic-polycytidylic acid (poly I:C), a synthetic analog of double-stranded RNA (dsRNA) known to be present in viral infection. A combination of bioactivity-guided isolation with an NMR-based identification led to the isolation of 4-acetylantroquinonol B (1), along with seven compounds. The structure of new compounds (4 and 5) was elucidated by spectroscopic experiments, including MS, IR, and NMR analysis. The anti-inflammatory activity of all isolated compounds was assessed at non-cytotoxic concentrations. 4-Acetylantroquinonol B (1) was the most potent compound against poly I:C-induced NO production in RAW 264.7 cells with an IC50 value of 0.57 ± 0.06 μM. [ABSTRACT FROM AUTHOR]

It has been reported that 4,5-dihydropyrazole and thiazole derivatives have many biological functions, especially in the aspect of anti-inflammation. According to the strategy of pharmacophore combination, we introduced thiazolinone and dihydropyrazole moiety into steroid skeleton to design and synthesize a novel series of D-ring substituted steroidal 4,5-dihydropyrazole thiazolinone derivatives, and assessed their in vitro anti-inflammatory profiles against Lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 macrophage cells. The anti-inflammatory activities assay demonstrated that compound 12e was considered as the most effective anti-inflammatory drug, which suppressed the expression of pro-inflammatory mediators including nitric oxide (NO), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α), it also dose-dependently inhibited the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in LPS-induced RAW 264.7 macrophage cells. Furthermore, the results of the Western blot analysis showed a correlation between the inhibition of the Nuclear factor-kappa B (NF-κB) and Mitogen-activated protein kinases (MAPKs) signaling pathways and the suppressive effects of compound 12e on pro-inflammatory cytokines. Molecular docking studies of compound 12e into the COX-2 protein receptor (PDB ID: 5IKQ) active site was performed to rationalize their COX-2 inhibitory potency. The results were found to be in line with the biological findings as they exerted more favorable interactions compared to that of dexamethasone (DXM), explaining their remarkable COX-2 inhibitory activity. The findings revealed that these candidates could be identified as potent anti-inflammatory agents, compound 12e could be a promising drug for the treatment of inflammatory diseases. [Display omitted] • This study provided a novel, efficient and simple synthesis of steroidal 4,5-dihydropyrazole thiazolinone derivatives. • In vitro activity assay screened compound 12e with excellent anti-inflammatory biological activity and low cytotoxicity. • 12e inhibited LPS-induced expression of the pro-inflammatory cytokines iNOS, COX-2, TNF-α and IL-6 on RAW 264.7 cells. • 12e exerted anti-inflammatory effects through down-regulation of NF-κB/MAPKs pathway. [ABSTRACT FROM AUTHOR]

Inflammation, an important biological protective response to tissue damage or microbial invasion, is considered to be an alarming signal for the progress of varied biological complications. Based on the previous reports in the literature that proved the noticeable efficacy of pyrazole and thiazole scaffold as well as nitrogen heterocyclic based compounds against acute and chronic inflammatory disease, a new set of novel D-ring substituted steroidal 4,5-dihydropyrazole thiazole derivatives were synthesized and evaluated their anti-inflammatory activities in vitro. Preliminary structure-activity relationship (SAR) analysis was conducted by their inhibitory activities against nitric oxide (NO) release in lipopolysaccharide (LPS)-induced RAW 264.7 cells, and the optimal compound 12b [3β-hydroxy-pregn-5-en-17β-yl-5′- (o- chlorophenyl)− 1′-(4′′- phenyl -[1′′, 3′′]- thiazol-2′′- yl) − 4′,5′-dihydro − 1′H-pyrazol − 3′- yl] exhibited more potent anti-inflammatory activity than the positive control treatment methylprednisolone (MPS), with an IC 50 value of 2.59 μM on NO production and low cytotoxicity against RAW 264.7 cells. In further mechanism study, our results showed that compound 12b significantly suppressed the production of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and inhibited the expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) through blocking NF-κB p65 nuclear translocation and phosphorylation of IκBα. Compound 12b also attenuated LPS-induced activation of c-Jun amino-terminal kinase (JNK) and p38 phosphorylation in RAW 264.7 cells. Molecular docking study revealed the strong binding affinity of compound 12b to the active site of the COX-2 proteins, which confirmed that compound 12b acted as an anti-inflammatory mediator. These results indicate that steroidal derivatives bearing 4,5-dihydropyrazole thiazole structure might be considered for further research and scaffold optimization in designing anti-inflammatory drugs and compound 12b might be a promising therapeutic anti-inflammatory drug candidate. [Display omitted] • This study provided a novel series of D-ring substituted steroidal 4,5-dihydropyrazole thiazole derivatives. • The structures of all targeted synthesized compounds had been characterized. • The anti-inflammatory biological activity of targeted synthesized compounds was assayed. • In vitro activity assay screened compound 12b with excellent anti-inflammatory biological activity in LPS-induced RAW 264.7 cells. [ABSTRACT FROM AUTHOR]

[Display omitted] • Tetrahydropiperic acid (THPA) can effectively attenuate the inflammatory response, both in-vitro and in-vivo suggesting, THPA as a potential novel anti-inflammatory drug candidate in the treatment of inflammatory disorders. • THPA significantly decreased the production of NO and pro-inflammatory cytokines (IL-6, IL-1β & TNF-α) in LPS stimulated RAW 264.7 macrophages and in-vivo experimental model. • THPA effectively decreased carrageenan-induced paw edema and showed protective effect in LPS challenged mice. • The anti-inflammatory potential of THPA is attributed to downregulation of NF-κB signaling pathway. The present study describes the synthesis, characterization, and evaluation of tetrahydropiperine (THP), piperic acid (PA), and tetrahydropiperic acid (THPA) as anti-inflammatory agents. THPA demonstrated potent anti-inflammatory activity among all the compounds. The anti-inflammatory potential was investigated in both in-vitro and in-vivo experimental models. Our findings demonstrated that THPA effectively suppressed the production of pro-inflammatory mediators, including nitric oxide and pro-inflammatory cytokines (TNF-α, IL-6, and IL-1β) in both in vitro and in vivo. Additionally, THPA attenuated the expression of i-NOS and COX-2 in RAW 264.7 macrophages. The oral administration of THPA significantly reduced carrageenan induced paw edema thickness and alleviated liver, lung, and kidney injury induced by LPS. THPA also reduced the infiltration of inflammatory cells, prevented the occurrence of significant lesions, and mitigated tissue damage. Moreover, THPA significantly improved the survival rate of mice challenged with LPS. Our western blot studies also found that LPS induced NF-κB activation was downregulated by treatment with THPA in an in vivo system. These results collectively illustrated the potential of THPA as a therapeutic agent for treating inflammatory diseases. [ABSTRACT FROM AUTHOR]

Purpose: To investigate the potential anti-inflammatory effects of the seeds of Opuntina humifusa and its active constituents. Methods: The extract of O. humifusa seeds was tested for the inhibition of nitric oxide (NO) production in liposaccharide (LPS)-stimulated RAW 264.7 cells using Griess reagent. The active constituents were isolated using bioassay-guided isolation methods. The effects of the active constituent on NO, proinflammatory cytokines, nuclear factor-kappa B (NF-κB) and nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor (IκB) were evaluated by enzyme-linked immunosorbent assay (ELISA) and western blot analysis. Results: The seed extract of O. humifusa significantly attenuated LPS-induced NO production in RAW 264.7 cells (p < 0.05). Bioassay-guided fractionation resulted in the isolation of isoamericanin A as an active constituent. Isoamericanin A reduced LPS-induced production of NO, iNOS, and proinflammatory cytokines (TNF-α and IL-6) in a concentration-dependent manner (p < 0.05). Furthermore, the effect was accompanied by decreased translocation of NF-κB from the cytosol to the nucleus and the decreased phosphorylation of IκB in the cytosol induced by LPS (p < 0.05). Conclusion: The seed extract of O. humifusa and its active constituent, isoamericanin A, have antiinflammatory effects in LPS-stimulated RAW 264.7 cells, suggesting that they have potentials as antiinflammatory agents. [ABSTRACT FROM AUTHOR]

Danshensu, isolated from Salvia miltiorrhiza (Danshen), is known to have anti-inflammatory properties. Therefore danshen is extensively used in many nutraceutical formulations. Reactive oxygen species are essential for the development of hypoxia-induced inflammation. Generation of reactive oxygen species by infiltrating macrophages is common in various diseases such as cardiovascular disease, neurodegenerative disease, tumor, and aging. To explore the mechanism underlying the attenuation of inflammation, we used RAW 264.7 cells as a model and hypoxia as an inducer of inflammation. The results showed the protective mechanism of danshensu on reactive oxygen species production, hypoxia-inducible factor 1-alpha expression, c-Jun N-terminal kinase phosphorylation, nuclear translocation of nuclear factor kappa B, and inducible nitric oxide synthase expression following hypoxia in RAW 264.7 cells. [ABSTRACT FROM AUTHOR]

• A new family of D-ring pyrazoline derivatives of pregn-5-enes were synthesized. • The structures of all targeted synthesized compounds had been characterized. • The anti-inflammatory biological activity of synthesized compounds was assayed. • Compound 4g was with excellent anti-inflammatory activity and low cytotoxicity. To identify new potential anti-inflammatory agents, a number of novel steroidal derivatives with nitrogen heterocyclic side chains 4a-4l were synthesized and evaluated for their anti-inflammatory effects in activated RAW 264.7 macrophage cells. The synthesis scheme involves two steps, Claisen-Schmidt condensation with the corresponding pregnenolone and aromatic aldehydes as the first step followed by nucleophilic addition of thiosemicarbazide across an α, β-unsaturated carbonyl as a later step. Compound structures were confirmed by 1H NMR, 13C NMR, HRMS, and IR. The compounds were assayed to test their anti-inflammatory effects in activated RAW 264.7 cells. Compound 4g , 3β-hydroxy-pregn-5-en-17β-yl-5′-(m-fluorophenyl)-4′, 5′-dihydro-1′-carbothioic acid amido pyrazole, was identified as the most potent anti-inflammatory agent of the analysed compounds, with an IC 50 value of 0.86 µM on nitric oxide (NO) production in lipopolysaccharide (LPS)-induced RAW 264.7 cells for 24 h compared to dexamethasone (IC 50 = 0.62 µM) and low cytotoxicity against RAW 264.7 cells. Compound 4g significantly inhibited NO produced by LPS-induced RAW 264.7 cells. Further studies showed that compound 4g markedly inhibited the expression of pro-inflammatory factors, including inducible nitric oxide synthase (iNOS), interleukin-6 (IL-6), tumour necrosis factor-α (TNF-α), and cyclooxygenase-2 (COX-2) in LPS-induced RAW 264.7 cells. These results indicate that derivatives bearing pyrazoline structure might be considered for further research and scaffold optimization in designing anti-inflammatory drugs and compound 4g might be a promising therapeutic anti-inflammatory drug candidate. [ABSTRACT FROM AUTHOR]

Background: There have been anecdotal reports from Thai hyperuricemic patients that the leaves of Lysiphyllum strychnifolium could reduce plasma uric acid level and relieve inflammation of gout. However, no research to support these effects has been conducted. Objectives: This study was aimed to evaluate the anti-inflammatory and hypouricemic effects of L. strychnifolium leaves extract and to investigate the pharmacological mechanisms of these effects. Materials and Methods: The anti-inflammatory effect of L. strychnifolium was evaluated in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Effects of L. strychnifolium on xanthine oxidase (XO) were examined in vitro and in vivo using potassium oxonate (PO)-induced hyperuricemic mice. In addition, the antioxidant activity of L. strychnifolium was determined. Results: L. strychnifolium significantly reduced the mRNA expression of cyclooxygenase-II, inducible nitric oxide synthase, transforming growth factor-β, and tumor necrosis factor-α in LPS-stimulated RAW 264.7 cells (P < 0.05). It exhibited a noncompetitive inhibition of XO activity with IC50and K i of 231 μg/ml and 177 μg/ml, respectively. Oral administration of L. strychnifolium (100 and 200 mg/kg) significantly lowered the plasma uric concentration (P < 0.05) in PO-induced hyperuricemic mice and inhibited 56.9% and 66.3% of the hepatic XO activity, respectively, compared to control hyperuricemic mice (P < 0.05). L. strychnifolium did not significantly decrease the protein expression of solute carrier family 22 member 12 in the renal cortex. Total phenolic and flavonoid contents were determined to be 197.8 ± 5.8 mg gallic acid equivalence/g extract and 32.2 ± 1.2 mg quercetin equivalent/g extract, respectively. The IC50of the inhibition of 2,2-diphenyl-1-picrylhydrazyl radical was 44.32 μg/ml. Conclusion: The present study first provided scientific evidence for the anti-inflammatory, anti-hyperuricemic and antioxidant effects of L. strychnifolium leaves extract in vitro and in vivo, suggesting the possibility of this plant to treat gout. Abbreviations used: ABTS: 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid); COX-II: Cyclooxygenase-II; DMEM: Dulbecco's modified Eagle medium; DPPH: 2,2-Diphenyl-1-picrylhydrazyl; FBS: Fetal bovine serum; FRAP: Ferric reducing antioxidant power; GAE: Gallic acid equivalence; GLUT9: Glucose transporter 9; iNOS: Inducible nitric oxide synthase; LPS: Lipopolysaccharide; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide; PO: Potassium oxonate; P/S: Penicillin/streptomycin; QE: Quercetin equivalent; SLC22A12: Solute carrier family 22 member 12; TNF-α: Tumor necrosis factor-α; TGF-β: Transforming growth factor-β; URAT: Urate-anion transporter 1; XO: Xanthine oxidase. [ABSTRACT FROM AUTHOR]

Context: Chatuphalatika (CTPT), is a Thai herbal formulation mixture of Phyllanthus emblica Linn. (Euphorbiaceae), Terminalia belerica Linn. (Combretaceae), T. chebula and the fruit of T. arjuna (Roxb.) Wight & Arn. CTPT is considered to exert anti-inflammatory and antihyperuricemic effects, but there have been no reports to demonstrate these pharmacological effects in a quantitative manner. Objectives: To investigate the antioxidative, anti-inflammatory and antihyperuricemic effects of CTPT. Materials and methods: Antioxidant activities of CTPT extracts were measured in vitro by DPPH, ABTS and FRAP assays, and anti-inflammatory effect by measuring inflammatory mediator production induced by lipopolysaccharide (LPS) in RAW264.7 macrophages. The mechanism of the hypouricemic effect was investigated using oxonate-induced hyperuricemic ddY mice treated with oral administrations of CTPT at 250, 500 and 1000 mg/kg. Results: Antioxidant activities of CTPT measured by ABTS and FRAP assays were 1.35 g TEAC/g extract and 10.3 mmol/100 g extract, respectively. IC50 for the inhibition of DPPH radical was 13.8 µg/mL. CTPT (10 µg/mL) significantly downregulated the mRNA expression of TNF-α and iNOS in RAW 264.7 cells. Lineweaver-Burk analysis of the enzyme kinetics showed that CTPT inhibited xanthine oxidase (XOD) activity in a noncompetitive manner with the Ki of 576.9 µg/mL. Oral administration of CTPT (1000 mg/kg) significantly suppressed uric acid production by inhibiting hepatic XOD activity, and decreased plasma uric acid levels in hyperuricemic mice by approximately 40% (p < 0.05). Conclusions: This study demonstrated for the first time the antioxidative, anti-inflammatory and antihyperuricemic effects of CTPT in vivo and in vitro, suggesting a possibility of using CTPT for the treatment of hyperuricemia in gout. [ABSTRACT FROM AUTHOR]

Ginsenoside Rh2, a protopanaxadiol saponin from ginseng, has been reported to have strong anti-inflammatory activity. However, the concentration of ginsenoside Rh2 is very low (>0.001%) in the total ginseng extracted, which is not enough for production despite its high pharmacological effects. Thus, in this study, we evaluated the anti-inflammatory effect of ginsenoside Rh2-mix (GRh2-mix) on lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophage cells. From the high-performance liquid chromatography analysis, it was confirmed that the GRh2-mix was mainly composed of 20(S)-Rh2, 20(R)-Rh2, Rk2, and Rh3. The LPS-stimulated RAW 264.7 cells were treated with different concentrations of GRh2-mix (100, 200, 400, 500 μg/mL). The cell counting kit-8 assay showed that the GRh2-mix treatment increased cell proliferation in LPS-stimulated RAW 264.7 murine macrophage cells. The GRh2-mix inhibited nitric oxide production in a dose-dependent manner, suggesting an anti-inflammatory effect. Furthermore, reverse transcription polymerase chain reaction and Western blot results also indicated that the GRh2-mix suppressed inflammatory genes such as iNOS, TNF-α, COX-2, IL-1β, IL-6, and NF-κB. In summary, these results suggest that the GRh2-mix exhibits anti-inflammatory activity via the downregulation of the NF-κB pathway and has high efficiency with a simple production procedure. [ABSTRACT FROM AUTHOR]

Objective(s): In traditional Chinese medicine, gamboge can detoxify bodies, kill parasites, and act as a hemostatic agent. Recent studies have demonstrated that gambogic acid (GBA) suppressed inflammation in arthritis, and also presented antitumor effect. Thus, this study investigated the new biological properties of GBA on macrophages. Materials and Methods: RAW 264.7 cells were pretreated with GBA at different concentrations (10, 20, 40, 80, 160, 320 nM) for 24 hrs, and then cell viability was measured using Cell Counting Kit (CCK)-8 assays. Pro-inflammatory cytokines such as TNF-α, IL-6 and IL-1α were determined using ELISA kits and qPCR. Then nitrite concentration was calculated according to a standard curve. At last, the effect of GBA on MAPK and NF-κB signaling pathways was assessed by western blot and luciferase reporter gene assay. Results: GBA (IC50: 260 nM) suppressed the TNF-α, IL-6 and IL-1β expression induced by lipopolysaccharide (LPS) in RAW 264.7 cells. The expression of TNF-α, IL-6 and IL-1&#; decreased to 30-50% and 70-75% in the high-dose (160 nM) and low-dose (40 and 80 nM) GBA groups, respectively. Furthermore, the nitric oxide (NO) production and the activation of NF-κB, ERK, and JNK pathways were significantly reduced in high-dose (160 nM) GBA only, while p38 pathway was inhibited at both low (40 and 80 nM) and high (160 nM) concentration of GBA. Conclusion: These data suggested that GBA inhibited LPS-induced production of pro-inflammatory cytokines including TNF-α, IL-6 and IL-1β mainly through the suppression of the p38 pathway. [ABSTRACT FROM AUTHOR]

Background: Panax ginseng has been used as a traditional medicine for various pathological conditions. Ginsenoyne C (GSC) is a polyacetylene, minor constituent. Objective: The objective of the present study was to determine the anti-inflammatory activities of GSC for the protection of inflammation. Materials and Methods: The effects of GSC on cell viability and nitric oxide (NO) production in lipopolysaccharide (LPS)–induced RAW 264.7 cells were investigated through MTT assay and the Griess reaction, respectively. The levels of inflammatory cytokines such as interleukin (IL-1β) and IL-6 and inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), and pERK were measured using enzyme-linked immunosorbent assay and Western blot analysis, respectively. Results: GSC suppressed NO production in LPS-induced RAW 264.7 cells. GSC suppressed the production of inflammatory mediators through downregulating phosphorylation of extracellular regulated kinases signaling in LPS-induced RAW 264.7. Conclusion: GSC has a potential therapeutic agent for protection of inflammation. Abbreviation used: CCK: Cell counting kit; COX-2: Cyclooxygenase-2; ELISA: Enzyme linked-immunosorbent assay; ERK: Extracellular regulated kinases; FBS: Fetal bovine serum; FEG: Fermented ginseng extract; GSC: Ginsenoyne C; IL: Interleukin; iNOS: Inducible nitric oxide synthase; LPS: Lipopolysaccharide; MAPK: Mitogen-activated protein kinases; NO: Nitric oxide; PVDF: Polyvinylidene difluoride; SDS-PAGE: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis [ABSTRACT FROM AUTHOR]

Ethnopharmocological relevance Cortex Eucommiae (CE), the bark of Eucommia ulmoides Oliv., has been traditionally used for its kidney-tonifying and bone- and tendon-enhancing properties in Korea, China, and Japan. CE has been historically prescribed for inflammatory conditions such as arthritis of the knee and ankle. Aim of the study Although CE has recently been shown to suppress inflammation in scientific studies, whether this effect involves modulation of the toll-like receptor 4 (TLR-4) pathway is currently unknown. Materials and methods The modulatory effect of CE on the TLR-4 pathway, both myeloid differentiation primary response gene 88 (Myd88)-dependent and independent, was investigated through real-time reverse transcriptase-polymerase chain reaction (RT-PCR), western blotting, and a reporter gene assay in lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophages. Results CE dose-dependently inhibited nitric oxide production without significant cytotoxicity with an IC 50 of 356.23 μg/mL. In addition, CE down-regulated both LPS-induced mRNA and protein expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) in a dose-dependent manner. CE suppressed LPS-induced activation of nuclear factor-κB (NF-κB) and the mitogen-activated protein kinase (MAPK) pathways, which together comprise the Myd88-dependent TLR-4 pathway. The phosphoinositide 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) pathway was also down-regulated by CE in a dose-dependent manner. CE additionally suppressed LPS-induced activation of interferon-β (IFN-β) and signal transducer and activator of transcription (STAT) pathway, which is associated with the Myd88-independent TLR-4 pathway. Conclusions CE down-regulated both Myd88-dependent and independent TLR-4 pathways, thus exerting anti-inflammatory effects. These results suggest that CE may be used as a potential therapeutic agent against chronic inflammatory diseases. [ABSTRACT FROM AUTHOR]

Natural borneol (NB, called "Bingpian") is an important traditional Chinese medicine to restore consciousness, remove heat and relieve pain, all of which are inflammation-related diseases. Recently, due to the limited source of NB, synthetic borneol (SB) is widely used as a substitute for NB in clinics. However, little is known about the effects of SB instead of NB. Herein, the aim of the present study was to compare NB and SB on chemical profiles by gas chromatography-mass spectrometer (GC-MS) analysis, anti-inflammatory activity in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages, and ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) metabolomic approaches in endotoxic fever induced in rats. Results showed that, in total, 13 volatile components could be identified in NB and SB by GC-MS analysis, in which a significant difference between them still existed. The main constituents in SB were iso-borneol and borneol, while borneol contributes to 98.96% of the amount in NB. Additionally, both NB and SB exhibited remarkable anti-inflammatory effects to reduce the level of inflammatory factors including NO, TNF-α and IL-6 in LPS-induced RAW 264.7 macrophages, and lower the high body temperature in rats with endotoxic fever induced by LPS. Moreover, it seems that NB exhibited higher efficacy than SB. The unequal bioactive efficiency between NB and SB was also indicated by means of non-targeting metabolomics. Based on UPLC-Q-TOF/MS technology, 12 biomarkers in the serum of fever rats were identified. Pathway analysis revealed that the anti-fever effect of NB and SB was related to regulating the abnormal glycerophospholipid, linoleic acid and alpha-linoleic acid metabolism pathways in the fever model. Results indicated that there was still a great difference between NB and SB involving chemical constituents, anti-inflammation activity and the ability to regulate the abnormal metabolism pathways of the fever model. Certainly, further studies are warranted to better understand the replacement rationale in medicinal application. [ABSTRACT FROM AUTHOR]

Objective: Opuntia humifusa (O. humifusa) Raf. has been used for the prevention and treatment of rheumatoid arthritis, inflammation, and cancer. Our study was designed to unveil the anti-nociceptive and anti-inflammatory effects of the methanolic extract of O. humifusa Raf stem (OHS). Materials and Methods: The anti-nociceptive effect was measured by hot plate, acetic acid-induced writhing, and tail flick assays in mice and rats. Moreover, the anti-inflammatory effect was measured by vascular permeability and carrageenan and serotonin-induced paw edema tests in rats. Furthermore, antiinflammatory effect was also measured using macrophage-like LPS-induced RAW 264.7 cells. Results: OHS extract inhibited acetic acid-induced writhing (p<0.0001), and delayed the reaction time of mice to the hot plateinduced thermal stimulation (p<0.0001) and tail flick tests (p<0.05). OHS extract attenuated the carrageenan and serotonininduced paw edema in rats (p<0.001). Similarly, OHS extract significantly decreased Evans blue concentration in acetic acid induced vascular permeability test (p<0.0001), revealing its strong anti-inflammatory effect. Finally, among four different fractions of OHS extract, n-butanol fraction strongly decreased NO production (p<0.0001) and iNOS expression in LPS-induced RAW 264.7 cells. Conclusion: Our results suggest that the methanolic extract of O. humifusa stem can be used to develop a therapeutic or supportive drug and/or functional food against pain and inflammation related diseases. [ABSTRACT FROM AUTHOR]

Protein interacting with C kinase 1 (PICK1) is a scaffolding protein mainly implicated in neurological diseases, however, the function of PICK1 in acute liver injury (ALI) remains unknown. Our study found a dramatical decrease in mRNA and protein levels of PICK1 in liver tissues and isolated Kupffer cells (KCs) from the liver in mice with ALI. Furthermore, pretreatment the mice with ALI with FSC-231, a pharmacological inhibitor of PICK1, could significantly augment inflammatory response. Furthermore, in vitro studies showed that both lipopolysaccharide (LPS) and interferon gamma (IFN-γ) significantly reduced the expression of PICK1, while IL-4 elevated its expression in RAW 264.7 cells. Additionally, over-expression of PICK1 inhibited the expression of M1 biomarkers by suppressing NF-κB activity, and enhanced the expression of M2 biomarkers by promoting STAT6 activity. In contrast, knockdown of PICK1 or FSC-231 pretreatment promoted M1 polarization and suppressed M2 polarization. Besides, caveolin-1 was identified as a potential target gene controlled by PICK1 in RAW 264.7 cells. Mechanistic investigation revealed a dual role of PICK1 in regulating macrophage polarization and implied PICK1 as a potential therapeutic target in ALI. [ABSTRACT FROM AUTHOR]

Kaempferol 3- O -(2G-glucosylrutinoside)-7- O -glucoside (KGG) has isolated from Hosta plantaginea flowers and possessed an inhibitory effect on cyclooxygenase 2 (COX-2), could be effective in inhibiting inflammation. However, the anti-inflammatory activity and mechanism of KGG remain unknown. In this study, for the first time, the anti-inflammatory effect of KGG and its potential molecular mechanisms were explored in cells. KGG had no cytotoxicity at concentrations of 1.25, 2.5, 5, 10, 20, and 40 μM by Cell Counting kit-8 assay in RAW 264.7 cells. Besides, KGG concentration-dependently (1.25, 2.5, and 5 μM) inhibited secretions of nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β), and IL-6 in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Western blot showed that the phosphorylation of nuclear factor kappa-B (NF-κB) p65, inhibitor of NF-κB (IκB), p38 MAPK, c-Jun N-terminal kinases (JNK), extracellular signal-regulated kinase (Erk), and protein kinase B (Akt), together with inducible nitric oxide synthase (iNOS) and COX-2 were significantly attenuated by KGG (1.25, 2.5, and 5 μM) in a concentration-dependent relationship. Meanwhile, KGG remarkably enhanced the protein expression of IκB. Taken together, KGG may be one of bioactive phytochemicals from H. plantaginea flowers, and be an anti-inflammatory agent via inhibiting NF-κB, mitogen-activated protein kinases (MAPKs), and Akt signaling pathways. [Display omitted] • Kaempferol 3- O -(2G-glucosylrutinoside)-7- O -glucoside (KGG) exerts anti-inflammatory effect through blocking NF-κB, MAPKS, and Akt pathways. • KGG might be a promising candidate against inflammation. [ABSTRACT FROM AUTHOR]

Geraniin, a major polyphenolic compound of Geranium sibiricum L, has long been used as an important Chinese herbal medicine for the treatment of a variety of inflammatory pathologies. However, the underlying anti-inflammatory molecular mechanisms of this compound are not clear. The aim of the present study was to investigate the anti-inflammatory activities of geraniin and elucidate the underlying mechanisms. The anti-inflammatory effects of geraniin were studied by using lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Geraniin suppressed the inducible nitric oxide synthase (iNOS) expression, and inhibited reactive oxygen species (ROS) production. Subsequent studies demonstrated that geraniin effectively reduced production of NO and pro-inflammatory cytokines. These effects were mediated by impaired translocation of nuclear factor (NF)-κB and inhibition of the phosphorylation of Akt in LPS-stimulated RAW 264.7 cells. Furthermore, geraniin induced heme oxygenase-1 (HO-1) expression via activation of transcription factor Nrf2. This study gives scientific evidence that geraniin inhibits the LPS-induced expression of inflammatory mediators via suppression of Akt-mediated NF-κB pathway as well as up-regulation of Nrf2/HO-1 pathway, indicating that geraniin has a potential application in inflammatory conditions. [ABSTRACT FROM AUTHOR]

Three LAB strains (Pediococctis acidilactici PA320, Lactobacillus plantarum LP109 and L. platitarum LP110) were selected for testing the acid, bile salt tolerance, and adhesion capacity in this study. Furthermore, the reduced capacity of allergy immune reaction was evaluated with cells and animals. For cell tests, the three LAB strains were utilized for stimulating the macrophages RAW 264.7 and human peripheral blood mononuclear cells (PBMC). Enzyme-linked immunosorbent assay (ELISA) was applied to analyzing the cytokine generated from the simulation of LAB, such as Interferon-γ (IFN-γ), Interleukin-12 (IL-12), Interleukin-4 (IL-4), and Interleukin-10 (IL-10), to evaluate the function of LAB's anti-allergy effects. For assessing anti-allergic effects of LAB in vivo, murine model of atopic dermatitis was used. In vitro results showed the overall LAB strains enhanced Lhl cytokine secretion while the role in inhibiting Th2 cytokine secretion. The supplementation of LAB strains caused a reduction in IgE production upon ovalbumin sensitization. Significant reduction in inflammatory cytokine secretion was observed. Histological data showed that the LAB reduced ear thickness associated with skin allergy. In summary, antiallergy and anti- inflammatory effects of these three strains were clearly demonstrated in vitro and in vivo. [ABSTRACT FROM AUTHOR]

Lactobacillus species are potential probiotic bacteria for humans because of their capacity to improve certain biological functions in the host's immune system. In this study, we focused on three peptidoglycans (PGNs) derived from different Lactobacillus strains and investigated each PGN's anti-inflammatory capacity. Each PGN was analyzed using HPLC, MALDI–TOF/TOF MS and FTIR. All three PGNs displayed a β-1,4-linked N -acetylmuramic acid (MurNAc) and N -acetylglucosamine (GlcNAc) structure with some modifications in the polypeptides at the end of the MurNAc residue. In a new insight, we found that PGNs inhibit the release of inflammatory cytokines in LPS-induced RAW 264.7 cells; a capacity that may be related to the TLR-4 pathway. The goal for exploring PGN diversity in Lactobacillus strains is to better understand the potential use of Lactobacillus PGNs in food and pharmaceutical applications. [ABSTRACT FROM AUTHOR]

The article discusses a study which aims to observe the anti-inflammatory effect of cordycepin using lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Topics include the incubation and pretreatment of RAW 264.7 cells with cordycepin, adenosine, and LPS concentrations, determining the role of adenosine receptor to cordycepin, and exploring the association of cyclic adenosine monophosphate (cAMP) signalling in cordycepin's anti-inflammatory activity.

Here we aimed to investigate the anti-inflammatory effects of calcium citrate in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The anti-inflammatory effects of calcium citrate were investigated by assessing pro-inflammatory factors (NO, ROS, NF-κB, iNOS, and COX-2) and pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α). Treatment of cells with calcium citrate (10–100 μM) significantly reduced the generation of intracellular reactive oxygen species and increased the activities of the antioxidant enzymes superoxide dismutase, catalase, and glutathione peroxidase in LPS-stimulated macrophages. Calcium citrate was further shown to inhibit NO production in LPS-stimulated RAW 264.7 cells. The expression levels of iNOS, COX-2, and NF-κB were also suppressed by treatment with calcium citrate. Calcium citrate was furthermore found to significantly inhibit the production of IL-1β, IL-6, and TNF-α in response to LPS-stimulation. These findings demonstrate that calcium citrate may be an effective anti-inflammatory agent. [ABSTRACT FROM AUTHOR]

Abstract: Three sulfated phenolic compounds, juglanin B (11R)-O-sulfate (1), myricetin 3´-O-sulfate (2), and ampelopsin 3´-O-sulfate (3), were isolated from the leaves of Myrica rubra. Compound 1 was a new sulfated lignan, 2 was a new sulfated flavone, and 3 was a known sulfated flavone. The structures of the new compounds (1 and 2) were determined by acid hydrolysis and spectroscopic methods, including IR, FAB-MS, 1D and 2D NMR. The inhibitory activities of compounds 1–3 and their hydrolysates (1a–3a) against LPS-induced cytokine (TNF-α, IL-1β, and IL-6) production in macrophage RAW 264.7 cells were evaluated. The 2 new compounds (1 and 2) and their aglycones (1a and 2a) significantly reduced LPS-induced expression of iNOS and COX-2 proteins. [Copyright &y& Elsevier]

The young branches of C. cassia Blume (Cinnamomi Ramulus; Guizhi; 桂枝; C. cassia twigs) have long been used as an anti-pyretic, anti-rheumatic, anti-spasmodic and stomachic in traditional medicine. The aim of this study was to test the anti-inflammatory, anti-nociceptive, and anti-arthritic effects of Cinnamomum cassia twigs in acute and chronic arthritis rats. Subcutaneous injection of carrageenan for acute inflammation and complete Freund's adjuvant (CFA) for chronic arthritis was carried out in the hind paw of SD rats. The paw volume was measured by a plethysmometer; thermal hyperalgesia was tested using a thermal plantar tester; hyperalgesia was evaluated by ankle flexion evoked vocalizations. The c-Fos expression in the lumbar spinal cord was measured with the avidin-biotin-peroxidase technique. The nitric oxide (NO) generation in lipopolysaccharide (LPS)-induced RAW 264.7 cells was tested by Griess assay. An 80% ethanoic extract of the C. cassia twigs exhibited chronic anti-inflammatory and anti-arthritic activities by reducing the edema volume in the paws of CFA-induced chronic arthritis in rats. In addition, it showed analgesic effects through the recovery of the paw withdrawal latency stimulated by thermal hyperalgesia, and suppressing the vocalization scores evoked by ankle flexion in the hind paws of the arthritis rats. It also controlled c-Fos expression in the lumbar spinal cord of the arthritis rats. Moreover, the addition its 80%-ethanoic extract, specifically, its ethyl acetate fraction, powerfully suppressed the paw swelling in carrageenan-stimulated arthritis and the NO production in LPS-induced murine immune cells. C. cassia twigs may act as a viably sufficient therapeutic or preventive candidate for osteoarthritis and rheumatoid arthritis; additionally, it could prevent gastrointestinal damage with its gastric protection. [Display omitted] [ABSTRACT FROM AUTHOR]

The article focuses on the essential oils (KEO) of Lindera umbellata that can inhibit lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 cells, a mouse leukaemic monocyte macrophage cell line. It analyzes the anti-inflammatory effect of KEO and states that it suppresses LPS-induced pro-inflammatory cytokine production of nitric oxide (NO), interleukin-6 (IL-6), and tumor necrosis factor-alpha and can also be used as anti-inflammatory therapeutic products.

Purpose: Laminaria japonica is a representative marine brown alga used as a culinary item in East Asia. L. japonica extract was shown to exert various biological activities; however, its anti-inflammatory activity has not been reported. The aim of this study is to investigate the molecular mechanisms underlying its anti-inflammatory action. Methods: Anti-inflammatory mechanisms of L. japonica n-hexane fraction (LHF) were assessed using lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. An anti-inflammatory compound isolated from LHF by reverse-phase chromatography was identified using nuclear magnetic resonance (NMR) spectroscopy. Results: Our results indicate that LHF significantly inhibited LPS-stimulated nitric oxide (NO) and prostaglandin E (PGE) secretion in a dose-dependent manner and suppressed the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) with no cytotoxicity. As results, levels of pro-inflammatory cytokines were significantly reduced by pretreatment of LHF in LPS-stimulated RAW 264.7 cells. Treatment of LHF strongly suppressed nuclear factor-κB (NF-κB) promoter-driven expression and nuclear translocation of NF-κB by preventing proteolytic degradation of inhibitor of κB (IκB)-α in LPS-stimulated RAW 264.7 cells. Moreover, LHF inhibited the phosphorylation of Akt and mitogen-activated protein kinase (MAPK) in LPS-stimulated RAW 264.7 cells. One of the anti-inflammatory compounds was isolated from LHF and identified as fucoxanthin. Conclusions: These results indicate that the LHF-mediated inhibition of NO and PGE secretion in LPS-stimulated macrophages is regulated by NF-κB inactivation through inhibition of IκB-α, MAPKs, and Akt phosphorylation. LHF may be considered as a functional food candidate for the prevention or treatment of inflammatory diseases. [ABSTRACT FROM AUTHOR]

Abstract: Aims of the study: We recently reported the potential antioxidant and anti-inflammatory activities of umbelliferone 6-carboxylic acid (UMC) isolated from the whole plants of Angelica decursiva. In this study, we elucidated the anti-inflammatory mechanisms of UMC in vitro and in vivo. Methods: The inhibitory effects of UMC on the production of nitric oxide (NO), prostaglandin E2 (PGE2), and tumor necrosis factor-α (TNF-α), the expression of nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), the activation of nuclear factor kappa B (NF-κB) were evaluated using lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. The reactive oxygen species (ROS) generation inhibitory activity of UMC was evaluated using t-butyl hydroperoxide (t-BHP)-induced RAW 264.7 cells. Furthermore, the in vivo anti-inflammatory activity of UMC was evaluated using carrageenan induced mouse paw edema model. Results: UMC dose-dependently inhibited NO and PGE2 production by down-regulating iNOS and COX-2 protein expression in LPS-stimulated RAW 264.7 macrophages. UMC also suppressed the production of the proinflammatory cytokine TNF-α in LPS stimulated RAW 264.7 cells in a concentration dependent manner. In addition, UMC dose-dependently prevented LPS-induced nuclear translocation of NF-κB in RAW 264.7 macrophages. Furthermore, UMC exhibited the inhibitory activity against t-BHP-induced ROS generation in RAW 264.7 cells with an IC50 value of 705.1μg/ml. Moreover, UMC inhibited λ-carrageenan induced mouse paw edema by 70.40 and 60.20% at doses of 50 and 25mg/kg body weight, respectively. Conclusion: The combined results of this study indicate that UMC is an important anti-inflammatory constituent of A. decursiva and its anti-inflammatory effect was due to its ability to inhibit the production of inflammatory mediators via inhibition of NF-κB activation pathway. [Copyright &y& Elsevier]

Strong anti-inflammatory activity has been found in Laminaria japonica dichloromethane fraction (LDF); however, the molecular mechanisms underlying its anti-inflammatory activity are not reported. Our results indicated that LDF inhibited LPS-induced nitric oxide and prostaglandin E production in a dose-dependent manner and suppressed the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) in RAW 264.7 cells. Also, levels of pro-inflammatory cytokines such as tumor necrosis factor-α, interleukin (IL)-1β and IL-6 were remarkably reduced by LDF in LPS-treated RAW 264.7 cells. LDF greatly inhibited promoter activity of nuclear factor-κB (NF-κB) and translocation of NF-κB subunits by prevention of the degradation of inhibitor κB-α in LPS-treated RAW 264.7 cells ( p < 0.05). Moreover, LDF inhibited activation of mitogen-activated protein kinases and AKT in LPS-treated RAW 264.7 cells. These results indicate that the LDF downregulates iNOS and COX-2 expressions through the suppression of NF-κB pathway associated with inhibition of multiple signaling proteins. [ABSTRACT FROM AUTHOR]

Abstract: Ethnopharmacological relevance: Orostachys japonicus A. Berger (O. japonicus) is known to reduce the risk of many diseases. Aim of the study: We investigated the anti-inflammatory effects of the dichloromethane (DCM) fraction from O. japonicus (OJD) in LPS-stimulated RAW 264.7 cells. Materials and methods: NO was measured using the Griess method. Key pro-inflammatory cytokines and mediators including IL-1β, TLR4, iNOS, and COX-2; 2 important pro-inflammatory transcription factors, NF-κB p65 and IκBα; and MAPKs such as ERK1/2, JNK, and p38 were analyzed by Western blotting. Results: OJD significantly inhibited NO production, IL-1β, TLR4, iNOS, and COX-2 expression in LPS-stimulated cells. Additionally, it inhibited LPS-induced NF-κB p65 activation via inhibition of IκBα phosphorylation. Furthermore, phosphorylation of p38 and JNK was suppressed by OJD in a dose-dependent manner in the LPS-stimulated RAW 264.7 cells. Conclusions: Our data suggest that OJD inhibits the inflammatory response via suppression of NF-κB activation and MAPK signaling. [Copyright &y& Elsevier]

Abstract: We have recently reported that phlorofucofuroeckol A isolated from the edible brown algae Ecklonia stolonifera showed potential antioxidative and anti-inflammatory properties in macrophage stimulated by LPS treatments. In this study, we further investigated the pharmacological characteristic of phlorofucofuroeckol A in regulations of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 through regulatory and signaling pathways using LPS-treated RAW 264.7 cells. Treatment with 20μM of phlorofucofuroeckol A significantly decreased levels of iNOS and COX-2 mRNA induced by LPS stimulation. As results, levels of pro-inflammatory cytokines such as interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α were significantly reduced by treatments of phlorofucofuroeckol A in LPS-stimulated RAW 264.7 cells. Phlorofucofuroeckol A inhibited promoter activities of inflammatory-mediators (iNOS and COX-2) and transcriptional factors (nuclear factor-κB, NF-κB, and AP-1) in LPS-treated RAW 264.7 cells. Moreover, phlorofucofuroeckol A inhibited activation of Akt and p38 MAPK in LPS-treated RAW 264.7 cells. These results indicate that the phlorofucofuroeckol A regulates iNOS and COX-2 expressions through the NF-κB-dependent transcriptional control associated with inhibition of multiple signaling proteins, suggesting potential candidates of phloroglucinol derivatives for treatments of inflammatory diseases. [Copyright &y& Elsevier]

We previously reported that the increment of carbohydrate content in the Viscozyme® L (Novozyme Corp., Oklahoma City, OK, USA) extract of Lactobacillus brevis-fermented Ecklonia cava affected the inhibition of nitric oxide (NO) production and that it might be related to the polysaccharide compound. However, there is no report of anti-inflammatory effects of the polysaccharide or its biological mechanism. Here, we investigated the anti-inflammatory effects of the polysaccharide and its biological mechanism in lipopolysaccharide (LPS)-activated RAW 264.7 cells. The polysaccharide isolated from the Viscozyme extract of L. brevis-fermented E. cava (VLFEP) dose-dependently decreased LPS-stimulated NO production without cytotoxicity. Also, VLFEP significantly decreased the production of prostaglandin E2 (PGE2) at the 100 μg/mL concentration. In addition, VLFEP dose-dependently decreased the protein and mRNA expressions of inducible NO synthase, whereas it slightly decreased those of cyclooxygenase 2 and only at the 100 μg/mL concentration. Moreover, VLEFP dose-dependently decreased the productions and/or mRNA expressions of tumor necrosis factor-α and interleukin-6, compared with those of LPS only-stimulated cells. In further experiments, VLFEP considerably reduced the phosphorylation and degradation of inhibitory κB as well as the translocation of nuclear transcription factor-κB (NF-κB) p65 into the nucleus, and its DNA binding was markedly induced by LPS stimulation. This study suggests that VLFEP exerts anti-inflammatory effects by down-regulating the production and expression of pro-inflammatory cytokines and mediators via inhibiting the NF-κB pathway in LPS-stimulated RAW 264.7 cells. [ABSTRACT FROM AUTHOR]

Abstract: Some antibiotics, including minocycline, have recently been reported to display immunomodulatory properties in addition to their antimicrobial activity. The use of a compound with both immunomodulatory and antibacterial properties could be very interesting in the treatment of inflammatory bowel disease (IBD), so the aim of our study was to evaluate the anti-inflammatory effect of minocycline in several experimental models of IBD. Firstly, the immunomodulatory activity of the antibiotic was tested in vitro using Caco-2 intestinal epithelial cells and RAW 264.7 macrophages; minocycline was able to inhibit IL-8 and nitrite production, respectively. In vivo studies were performed in trinitrobenzenesulfonic acid (TNBS)-induced rat colitis and dextran sodium sulfate (DSS)-induced mouse colitis. The results revealed that minocycline exerted an intestinal anti-inflammatory effect when administered as a curative treatment in the TNBS model, modulating both immune and microbiological parameters, being confirmed in the DSS model; whereas none of the other antibiotics tested (tetracycline and metronidazole) showed anti-inflammatory effect. However, minocycline administration before the colitis induction was not able to prevent the development of the intestinal inflammation, thus showing that only its antimicrobial activity is not enough for the anti-inflammatory effect. In conclusion, minocycline displays an anti-inflammatory effect on different models of rodent colitis which could be attributed to the association of its antibacterial and immunomodulatory properties. [Copyright &y& Elsevier]

This research was undertaken to find the in vitro inflammatory action of lansai A-D produced by Streptomyces sp. SUC1. We investigated the effects of lansai C not only on the formation of nitric oxide (NO), prostaglandin E2 (PGE2), tumour necrosis factor (TNF-α), interleukin (IL)-1α, IL-6 and IL-10, but also on inducible NO synthase and cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS)-induced murine macrophage RAW 264.7 cells. The data obtained were consistent with the modulation of inducible nitric oxide synthase enzyme expression. A similar fashion was also observed when LPS-induced PGE2 release and COX-2 expression were tested. The significant inhibitory effects were shown in concentration-dependent manners. In addition, lansai C also mildly but significantly reduced the formation of TNF-α. These findings support the application of lansai C as anti-inflammatory agent. [ABSTRACT FROM AUTHOR]

Abstract: Ethnopharmacological relevance: Taiuiá or tayuya (Cayaponia tayuya, Cucurbitaceae) is a climbing, lignified plant with a large swollen root that has traditionally been used as an anti-inflammatory and anti-rheumatic agent in the folk medicine of Brazil, Peru, and Colombia. The aim of the study: We have assayed the pharmacological properties of a flavonoid fraction obtained from the butanol extract of Cayaponia tayuya roots using two models of topical mouse ear oedema, paying special attention to its influence on the induction on pro-inflammatory enzymes and peptidic mediators. Material and methods: The in vivo experiments involved both the acute oedema induced by a single application of TPA and the subchronic inflammation brought on by repeated applications of TPA. The effects on the induction of pro-inflammatory enzymes and peptidic mediators in RAW 264.7 macrophages were analyzed with the aid of Western blot analysis. Results: The extract was identified as a mixture of flavonoids in which vicenin-2, spinosin, isovitexin, and a mixture of swertisin and isoswertisin were found. In acute TPA-induced oedema in mouse ears, the flavonoid-enriched fraction (at a dose of 0.5mg/ear) inhibited the oedema by 66% (4.2±0.6mg vs. 12.3±1.4mg, P <0.01) while in the subchronic model, the inhibition reached 37% at a dose of 0.5mg/ear×7 applications (7.5±0.6mg vs. 11.9±1.3mg, P <0.05). When assayed in vitro, the flavonoid showed no toxicity at 33.45μg/mL on RAW 264.7 macrophages. Although the nitric oxide production in these cells was moderately reduced (42%) at 33.45μg/mL, the flavonoid-enriched fraction had no effect on TNF-α production. In addition, at 22.30μg/mL, the test sample inhibited both iNOS and COX-2 expression by 98% and 49%, respectively. Conclusion: These results indicate that the anti-inflammatory activity of flavonoids from tayuya roots most likely stems from their inhibition of the induction of the enzymes COX-2 and iNOS. [Copyright &y& Elsevier]

Scurrula ferruginea (Jack) Danser (locally known as 'Dedalu' or 'dian nan ji sheng' in Malaysia and China) is a hemi-parasitic shrub that is widely used as herbal medicine to treat inflammation, rheumatism, and stroke. However, the scientific basis of its anti-inflammatory function and mechanism remain to be proven. Aim of the study : To evaluate the anti-inflammatory activity as well as the preliminary mechanism of S. ferruginea parasitizing on Tecoma stans. The anti-inflammatory capability of freeze-dried stem aqueous extract was assessed via inhibition of inflammatory cytokines interleukin- (IL-) 1 β , IL-6, IL-10, and tumor necrosis factor-alpha (TNF- α) production in lipopolysaccharide (LPS) and interferon- γ (IFN- γ) stimulated RAW 264.7 macrophages. The underlying anti-inflammatory mechanism was deciphered through reverse transcriptase and real time quantitative polymerase chain reactions (RT-PCR and qPCR) for inducible NO synthase (i NOS), cyclooxygenase-2 (COX-2), IL-1 β , and TNF- α mRNA expression. The results exhibited that aqueous extract of freeze-dried S. ferruginea stem sample concentration-dependently inhibited IL-1 β protein production along with the down regulation of i NOS and IL-1 β mRNA expression. Moreover, it significantly suppressed the protein release of IL-6 and IL-10 in a concentration-dependent manner. However, it slightly reduced TNF- α at higher sample concentration (250 μg/mL) without affecting the mRNA expression levels of COX-2 and TNF- α. This study suggests that S. ferruginea parasitizing on Tecoma stans exerted anti-inflammatory capability attributed to inhibition of i NOS and IL-1 β mRNA expression, NO creation, IL-1 β , IL-6, IL-10, and TNF- α protein production, indicating this plant might be a useful plant-derived candidate against inflammation. Image 1 • The anti-inflammatory effect in vitro of Scurrula ferruginea had been proven. • S. ferruginea inhibited NO, IL-1 β , IL-6, IL-10, and TNF- α productions. • Decrease of iNOS and IL-1β in LPS/IFN-γ induced RAW 264.7. [ABSTRACT FROM AUTHOR]

Selaginella tamariscina (S. tamariscina) (Beauv.) Spring (Selaginellaceae) has been used in oriental medicine for the treatment of dysmenorrhea, chronic hepatitis, hyperglycemia, amenorrhea, hematuria, prolapse of the anus and metrorrhagia. In the present study, we isolated two strong anti-inflammatory compounds, the biflavonoids hinokiflavone (H) and 7′-O-methyl hinokiflavone (mH), from S. tamariscina and examined their anti-inflammatory activities in lipopolysaccharide (LPS)-mediated murine macrophages (RAW 264.7) and colon epithelial cells (HT-29). H and mH suppressed the production of the inflammatory mediators nitric oxide (NO), interleukin (IL)-6, IL-8, and tumor-necrosis factor (TNF)-α, which are most highly activated in inflammatory bowel disease (IBD). In addition, Western blot analysis revealed that H and mH suppressed the LPS-induced expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2, and the activation of nuclear factor-κB (NF-κB) and extracellular regulated kinases (ERK) 1/2. These results suggest that H and mH are compounds having potent anti-inflammatory effects that could be used to treat such diseases as IBD. [ABSTRACT FROM AUTHOR]